unc-50 Antibody
Description
Introduction to UNC-50 Antibody
UNC-50 antibodies are immunological tools specifically designed to detect and study the UNC-50 protein, a conserved inner nuclear membrane RNA-binding protein involved in intracellular trafficking. The canonical human UNC-50 protein has a molecular weight of 30.4 kDa and is localized to the nucleus and Golgi apparatus . These antibodies are critical for applications such as Western blotting (WB), ELISA, and flow cytometry (FACS), enabling researchers to investigate UNC-50's roles in processes like receptor trafficking and cancer progression .
Key Attributes:
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Gene Symbol: UNC50
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Protein Family: Unc-50 family
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Functions:
Role in Hepatocellular Carcinoma (HCC)
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Upregulation in HCC: UNC50 expression is significantly elevated in 45.5% of HCC tissues compared to non-cancerous tissues .
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EGFR Pathway Regulation:
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UNC50 knockdown reduces mRNA levels of EGFR downstream targets (CCND1, EGF, MMP7) .
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Promotes G1/S cell cycle transition by enhancing EGFR cell surface localization, increasing responsiveness to EGF .
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Mechanism: UNC50 facilitates EGFR recycling to the plasma membrane without altering total EGFR levels (Fig. 5) .
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Trafficking of Nicotinic Receptors
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In C. elegans, UNC-50 ensures proper trafficking of levamisole-sensitive AChRs (Lev-AChRs) to neuromuscular junctions .
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Loss of Function: unc-50 mutants exhibit Lev-AChR degradation via lysosomes, impairing locomotion .
Table 1: Functional Impact of UNC50 in HCC
| Parameter | Effect of UNC50 Knockdown | Effect of UNC50 Overexpression |
|---|---|---|
| CCND1 mRNA Levels | ↓ 60% (p < 0.01) | ↑ 2.5-fold (p < 0.01) |
| Cell Cycle (G0/G1 Phase) | ↑ 15% (serum-free conditions) | ↓ 10% (with EGF stimulation) |
| Proliferation (MTT Assay) | No change (normal conditions) | ↑ 30% (with sustained EGF) |
Table 2: UNC-50 Antibody Validation
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research indicates that the transmembrane Golgi protein UNC-50 plays a crucial role in regulating nicotinic receptor trafficking. PMID: 17853888
Q&A
Here’s a structured collection of FAQs for academic researchers working with UNC-50 Antibody, organized by research complexity and grounded in scientific methodology:
Advanced Research Questions
How can researchers resolve contradictions in UNC-50 antibody reactivity across species?
| Species | Reported Reactivity | Validation Method |
|---|---|---|
| Human | Confirmed | siRNA knockdown + Western blot |
| Mouse | Predicted | Recombinant protein overexpression |
| Zebrafish | Predicted | CRISPR knockout models |
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Troubleshooting:
What strategies address low signal-to-noise ratios in UNC-50 immunohistochemistry?
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Optimization steps:
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Pre-treat tissue sections with antigen retrieval (e.g., citrate buffer, pH 6.0).
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Employ tyramide signal amplification for low-abundance targets.
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Validate with orthogonal methods (e.g., RNAscope for UNC-50 mRNA co-localization).
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How can UNC-50 antibody data be contextualized within RNA-binding protein networks?
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Systems biology approach:
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Conduct co-immunoprecipitation (Co-IP) with RNA pull-down assays to identify UNC-50-binding partners (e.g., spliceosome components).
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Integrate with transcriptomic datasets (e.g., CLIP-seq) to map RNA targets.
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Methodological Challenges and Solutions
How to differentiate UNC-50 isoforms in proteomic studies?
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LC-MS/MS workflow:
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Digest proteins with trypsin and analyze peptides covering residues 5–33 (antibody epitope region).
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Use parallel reaction monitoring (PRM) to quantify isoform-specific peptides.
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What controls are critical for UNC-50 functional studies in RNA processing assays?
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Essential controls:
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siRNA-resistant UNC-50 rescue constructs to confirm phenotype specificity.
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RNase-treated samples to distinguish RNA-dependent vs. independent interactions.
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Data Interpretation Frameworks
How to reconcile conflicting UNC-50 expression data across cancer types?
| Cancer Type | Reported UNC-50 Role | Key Confounder |
|---|---|---|
| Glioblastoma | Oncogenic | Tumor heterogeneity |
| Breast Cancer | Tumor-suppressive | Stromal contamination |
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Resolution:
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Apply single-cell RNA-seq to dissect tumor vs. microenvironment expression.
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Use laser-capture microdissection to isolate pure cell populations.
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